Mechanisms and regulation of extracellular vesicle traffic in the nervous system

神经系统细胞外囊泡运输的机制和调节

• 批准号: 10063578
• 负责人: Avital Adah Rodal
• 金额: $ 35.38万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-01 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10063578
• 关键词: Acute; Address; Alzheimer' s Disease; Animals; Autophagocytosis; Behavior; Biogenesis; Biological; Biology; Brain; Cells; Cellular Membrane; Chemicals; Communication; Complement; Complex; Coupled; Cultured Cells; Cytoplasm; Data; Degradation Pathway; Detection; Disease; Drosophila genus; Electron Microscopy; Equilibrium; Foundations; Gene Expression; Gene Expression Regulation; Genetic; Genetic Transcription; Goals; Guanosine Triphosphate Phosphohydrolases; Health; Human; Image; Imaging Techniques; Label; Lighting; Measurement; Measures; Mediating; Membrane; Membrane Protein Traffic; Methods; Microscopy; Modeling; Molecular; Nature; Nervous System Physiology; Nervous system structure; Neurodegenerative Disorders; Neurologic; Neuromuscular Junction; Neurons; Parkinson Disease; Pathologic; Physiological; Physiology; Presynaptic Terminals; Proteins; Recycling; Regulation; Research; Resolution; Rest; Role; Signal Transduction; Sorting - Cell Movement; Structure; Synapses; Synaptic Membranes; Synaptic plasticity; System; Testing; Tissues; Vesicle; cell type; experimental study; extracellular vesicles; in vivo; insight; live cell imaging; nervous system disorder; novel; presynaptic; reconstitution; response; targeted treatment; tool; trafficking; uptake; vesicular release

The goal of this project is to understand the molecular mechanisms of extracellular vesicle (EV) trafficking in the nervous system in vivo. EVs are small vesicles secreted from donor cells that can carry a diverse array of cargoes, and have recently come to the forefront as a novel mode of intercellular traffic and communication in the brain. EVs are thought to contribute to many human health conditions, including the spread of pathological proteins in neurodegenerative disease. However, because most studies of EVs are conducted with cells in culture, little is known about the release, uptake and fate of EVs in the diverse interacting cell types of the intact nervous system. We have developed a system in which to study traffic of endogenous neuronal EV cargoes in a living animal, using cutting edge genetic and cell biological tools available in Drosophila. Using this system, we discovered an unexpected role for synaptic periactive zone (PAZ) membrane remodeling machinery in EV cargo sorting, stability and release from axon terminals at the Drosophila larval neuromuscular junction. We also found that endogenous EV cargo release is dynamically regulated by neuronal activity. Finally, we made the surprising discovery that released EVs are partially protected from the target cell cytoplasm, suggesting the possibility that additional regulatory steps may govern their exposure and/or degradation upon internalization to recipient cells. Our proposed research will elucidate how cellular membrane traffic machinery controls the release, uptake, and fate of EV cargoes. To achieve these goals we will use advanced Drosophila genetics, live cell imaging techniques, structured illumination microscopy, electron microscopy, and tissue-specific detection and manipulation of EV cargoes in donor and recipient cells. Specifically, we propose: 1) To elucidate in vivo mechanisms of EV traffic and release by PAZ proteins. 2) to determine how activity regulates neuronal traffic of endogenous EV cargoes and 3) to determine the fate of NMJ EV cargoes, at rest and in response to neuronal activity. Given the conserved nature of synaptic membrane trafficking machinery, our findings and tools will lay the foundation for new insights into endogenous EV traffic in many aspects of nervous system function, including in human neurological disease.

该项目的目的是了解细胞外囊泡（EV）运输的分子机制 体内神经系统。电动汽车是从供体细胞中分泌的小囊泡，可以携带多种阵列 货物，最近作为一种新颖的细胞间交通和通信方式走到了最前沿 大脑。电动汽车被认为会导致许多人类健康状况，包括病理的传播 神经退行性疾病中的蛋白质。但是，由于大多数电动汽车研究是用细胞中的细胞进行的 文化，关于电动汽车在多种相互作用的细胞类型中的发行，吸收和命运知之甚少 完整的神经系统。我们已经开发了一个研究内源性神经元EV流量的系统 使用果蝇中有尖端遗传和细胞生物学工具的活动物中的货物。使用 该系统，我们发现突触周围区域（PAZ）膜重塑的意外作用 电动汽车货物排序，稳定性和从果蝇幼虫的轴突终端释放的机械 神经肌肉连接。我们还发现，内源性EV货物释放受到动态调节 神经元活性。最后，我们做出了一个令人惊讶的发现，即释放的电动汽车受到部分保护 靶细胞细胞质，表明其他调节步骤可以控制其暴露 和/或内部化对受体细胞的降解。我们提出的研究将阐明细胞 膜交通机械控制电动汽车货物的发行，吸收和命运。为了实现这些目标 将使用先进的果蝇遗传学，活细胞成像技术，结构化照明显微镜， 电子显微镜以及组织特异性检测以及对供体和受体细胞中EV货物的操纵。 具体而言，我们建议：1）阐明EV流量的体内机制并通过PAZ蛋白释放。 2）到 确定活动如何调节内源性EV货物的神经元流量和3）确定的命运 NMJ EV货物，静止并响应神经元活性。鉴于突触的保守性质 膜贩运机械，我们的发现和工具将为内生的新见解奠定基础 神经系统功能的许多方面的EV流量，包括人类神经疾病。